Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1993 May;61(5):1895–1899. doi: 10.1128/iai.61.5.1895-1899.1993

Production of a murine monoclonal antibody that recognizes an epitope specific to Coccidioides immitis antigen 2.

R A Cox 1, M J Dolan 1, D M Magee 1, J N Galgiani 1
PMCID: PMC280781  PMID: 7682998

Abstract

Antigen 2 (Ag2) has been implicated as a T-cell-reactive component of the pathogenic fungus Coccidioides immitis. We report the production of a murine monoclonal antibody (MAb) of the immunoglobulin G2a isotype that recognizes an epitope specific to C. immitis Ag2. This specificity was evidenced by the finding that the MAb did not recognize other antigens present in coccidioidin or spherulin and did not show reactivity with antigenic extracts from Histoplasma capsulatum or Blastomyces dermatitidis. The epitope was labile to enzymatic digestion with pronase but resistant to treatment with glycolytic enzymes and to periodate oxidation. This peptide epitope appears to require conformational structure on the basis that it was not recognized by the MAb in immunoblots of antigen that had been electrophoresed in polyacrylamide gels under denaturing, reducing conditions. Immunoaffinity chromatography of spherulin on columns containing the MAb established that the MAb was effective as a ligand for isolating Ag2 from heterogeneous extracts. The production of a MAb which recognizes an Ag2-specific epitope and its utility as a ligand for isolating Ag2 will provide a valuable reagent for studies of this immunologically important antigen.

Full text

PDF

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Cox R. A. Antigenic structure of Coccidioides immitis. Immunol Ser. 1989;47:133–170. [PubMed] [Google Scholar]
  2. Cox R. A., Britt L. A. Antigenic heterogeneity of an alkali-soluble, water-soluble cell wall extract of Coccidioides immitis. Infect Immun. 1985 Nov;50(2):365–369. doi: 10.1128/iai.50.2.365-369.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cox R. A., Britt L. A. Antigenic identity of biologically active antigens in coccidioidin and spherulin. Infect Immun. 1987 Nov;55(11):2590–2596. doi: 10.1128/iai.55.11.2590-2596.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cox R. A., Britt L. A. Isolation of a coccidioidin component that reacts with immunoglobulin M precipitin antibody. Infect Immun. 1986 Sep;53(3):449–453. doi: 10.1128/iai.53.3.449-453.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cox R. A., Brummer E., Lecara G. In vitro lymphocyte responses of coccidioidin skin test-positive and -negative persons to coccidioidin, spherulin, and a coccidioides cell wall antigen. Infect Immun. 1977 Mar;15(3):751–755. doi: 10.1128/iai.15.3.751-755.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cox R. A., Huppert M., Starr P., Britt L. A. Reactivity of alkali-soluble, water-soluble cell wall antigen of Coccidioides immitis with anti-Coccidioides immunoglobulin M precipitin antibody. Infect Immun. 1984 Feb;43(2):502–507. doi: 10.1128/iai.43.2.502-507.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cox R. A., Vivas J. R. Spectrum of in vivo and in vitro cell-mediated immune responses in coccidioidomycosis. Cell Immunol. 1977 Jun 1;31(1):130–141. doi: 10.1016/0008-8749(77)90012-0. [DOI] [PubMed] [Google Scholar]
  8. Dolan M. J., Cox R. A. Production and characterization of a monoclonal antibody to the complement fixation antigen of Coccidioides immitis. Infect Immun. 1991 Jun;59(6):2175–2180. doi: 10.1128/iai.59.6.2175-2180.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dolan M. J., Cox R. A., Williams V., Woolley S. Development and characterization of a monoclonal antibody against the tube precipitin antigen of Coccidioides immitis. Infect Immun. 1989 Apr;57(4):1035–1039. doi: 10.1128/iai.57.4.1035-1039.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dugger K. O., Galgiani J. N., Ampel N. M., Sun S. H., Magee D. M., Harrison J., Law J. H. An immunoreactive apoglycoprotein purified from Coccidioides immitis. Infect Immun. 1991 Jul;59(7):2245–2251. doi: 10.1128/iai.59.7.2245-2251.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Galgiani J. N., Sun S. H., Dugger K. O., Ampel N. M., Grace G. G., Harrison J., Wieden M. A. An arthroconidial-spherule antigen of Coccidioides immitis: differential expression during in vitro fungal development and evidence for humoral response in humans after infection or vaccination. Infect Immun. 1992 Jul;60(7):2627–2635. doi: 10.1128/iai.60.7.2627-2635.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Huppert M., Adler J. P., Rice E. H., Sun S. H. Common antigens among systemic disease fungi analyzed by two-dimensional immunoelectrophoresis. Infect Immun. 1979 Feb;23(2):479–485. doi: 10.1128/iai.23.2.479-485.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lecara G., Cox R. A., Simpson R. B. Coccidioides immitis vaccine: potential of an alkali-soluble, water-soluble cell wall antigen. Infect Immun. 1983 Jan;39(1):473–475. doi: 10.1128/iai.39.1.473-475.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mitchell C. G., Smith R., Lehner T. Recognition of carbohydrate and protein epitopes by monoclonal antibodies to a cell wall antigen from Streptococcus mutans. Infect Immun. 1987 Mar;55(3):810–815. doi: 10.1128/iai.55.3.810-815.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ward E. R., Jr, Cox R. A., Schmitt J. A., Jr, Huppert M., Sun S. H. Delayed-type hypersensitivity responses to a cell wall fraction of the mycelial phase of Coccidioides immitis. Infect Immun. 1975 Nov;12(5):1093–1097. doi: 10.1128/iai.12.5.1093-1097.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Woodward M. P., Young W. W., Jr, Bloodgood R. A. Detection of monoclonal antibodies specific for carbohydrate epitopes using periodate oxidation. J Immunol Methods. 1985 Apr 8;78(1):143–153. doi: 10.1016/0022-1759(85)90337-0. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES